Nanoscale spatio-temporal diffusion modes measured by simultaneous confocal and STED imaging
File(s)acs.nanolett.8b01190.pdf (3.66 MB)
Published version
Author(s)
Type
Journal Article
Abstract
The diffusion dynamics in the cellular plasma membrane provide crucial insights into molecular interactions, organization, and bioactivity. Beam-scanning fluorescence correlation spectroscopy combined with super-resolution stimulated emission depletion nanoscopy (scanning STED–FCS) measures such dynamics with high spatial and temporal resolution. It reveals nanoscale diffusion characteristics by measuring the molecular diffusion in conventional confocal mode and super-resolved STED mode sequentially for each pixel along the scanned line. However, to directly link the spatial and the temporal information, a method that simultaneously measures the diffusion in confocal and STED modes is needed. Here, to overcome this problem, we establish an advanced STED–FCS measurement method, line interleaved excitation scanning STED–FCS (LIESS–FCS), that discloses the molecular diffusion modes at different spatial positions with a single measurement. It relies on fast beam-scanning along a line with alternating laser illumination that yields, for each pixel, the apparent diffusion coefficients for two different observation spot sizes (conventional confocal and super-resolved STED). We demonstrate the potential of the LIESS–FCS approach with simulations and experiments on lipid diffusion in model and live cell plasma membranes. We also apply LIESS–FCS to investigate the spatiotemporal organization of glycosylphosphatidylinositol-anchored proteins in the plasma membrane of live cells, which, interestingly, show multiple diffusion modes at different spatial positions.
Date Issued
2018-06-12
Date Acceptance
2018-06-01
Citation
Nano Letters: a journal dedicated to nanoscience and nanotechnology, 2018, 18 (7), pp.4233-4240
ISSN
1530-6984
Publisher
American Chemical Society
Start Page
4233
End Page
4240
Journal / Book Title
Nano Letters: a journal dedicated to nanoscience and nanotechnology
Volume
18
Issue
7
Copyright Statement
© 2018 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY)
License, which permits unrestricted use, distribution and reproduction in any medium,
provided the author and source are cited.
License, which permits unrestricted use, distribution and reproduction in any medium,
provided the author and source are cited.
License URL
Sponsor
Marie Curie Career Integration Grant
Identifier
https://pubs.acs.org/doi/abs/10.1021/acs.nanolett.8b01190
Grant Number
PCIG13-GA-2013-618914
Subjects
Nanoscience & Nanotechnology
Publication Status
Published
Date Publish Online
2018-06-12